View source for Team:Queens Canada/Bacterial-Cell-Lysis


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<body><div><svg id="__SVG_SPRITE_NODE__" style="position: absolute; width: 0; height: 0" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink"></svg><div><div data-reactroot=""><div class="printable-container hide-print-links"><div><div class="protocolEditor"><div><!-- react-empty: 6 --><!-- react-empty: 7 --></div><div class="protocolEditor-pageContainer"><div class="protocolEditor-page"><div class="protocolEditor-contentWrapper"><h1 class="mediocre-titleEditor is-locked"><div class="editable selectable is-locked" contenteditable="false" data-allow-multiline="false" data-cid="pv884-title" data-index="0">Bacterial Cell Lysis</div></h1><div><h2>Introduction</h2><div class="intro-editor" data-editor="intro"><div class="editable selectable is-locked" contenteditable="false" data-allow-multiline="true" data-cid="pv884-intro" data-index="0">
</div></div><h2>Materials</h2><div class="materials-editor" data-editor="materials"><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><ul class="materials"><li class="material"><div class="material-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="m885" data-index="0">Resuspension buffer </div><ul class="components"><li class="component"><div class="component-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="c886" data-index="0,0">20 mM Tris-HCl (pH 7.6)</div></li><li class="component"><div class="component-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="c887" data-index="0,1">2 M Urea</div></li><li class="component"><div class="component-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="c888" data-index="0,2"></div></li></ul></li></ul></div><h2>Procedure</h2><div class="stepsEditor mediocre"><div class="editor" data-editor="steps"><div class="hiddenFocusEditable js-focusBearer" contenteditable="true" tabindex="-1"></div><div class="mediocre-item is-header" data-scroll-id="s889"><ul class="headers"><li><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s889" data-index="0"></div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s889" data-index="0" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ul></div><div class="mediocre-item is-step" data-scroll-id="s890"><ol class="steps"><li value="1"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s890" data-index="1">Harvest cells at 5 000 rpm for 20 minutes. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s890" data-index="1" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s891"><ol class="steps"><li value="2"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s891" data-index="2">Resuspend pellets in 25 mL of resuspension buffer.</div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s891" data-index="2" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s892"><ol class="steps"><li value="3"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s892" data-index="3">Sonicate to lyse cells.</div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s892" data-index="3" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s893"><ol class="steps"><li value="4"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s893" data-index="4">Sonicated pellets shoud be centrifuged at 12000 g for 20 min. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s893" data-index="4" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s894"><ol class="steps"><li value="5"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s894" data-index="5">After removing supernatant, pellets should be resuspended. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s894" data-index="5" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s895"><ol class="steps"><li value="6"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s895" data-index="6">Sonicate cells in 20 mL of Resuspension buffer (20 mM Tris-HCl (pH 7.6) and 6M urea). </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s895" data-index="6" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s896"><ol class="steps"><li value="7"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s896" data-index="7">Repeat twice.</div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s896" data-index="7" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s897"><ol class="steps"><li value="8"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s897" data-index="8">Keep supernatant after final centrifugation .</div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s897" data-index="8" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s898"><ol class="steps"><li value="9"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s898" data-index="9">Final supernatant was mixed with Ni-NTA agarose pre-equilibrated with resuspension buffer 2.</div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s898" data-index="9" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s899"><ol class="steps"><li value="10"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s899" data-index="10">Incubated for 1 hr at 4C. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s899" data-index="10" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s900"><ol class="steps"><li value="11"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s900" data-index="11">Pour entire mixture into column and initial flow through was collected for further analysis. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s900" data-index="11" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s901"><ol class="steps"><li value="12"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s901" data-index="12">Wash column with 5mL of wash buffer (20 mM Tris-HCl (pH 7.6), 6 M Urea, 20 mM imidazole) three times. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s901" data-index="12" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s902"><ol class="steps"><li value="13"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s902" data-index="13">Elute protein of interest with 1.5 mL of elution buffer (20 m Tris-HCl (pH 7.6), 6 M Urea, 0.3 M imidazole). </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s902" data-index="13" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div><div class="mediocre-item is-step" data-scroll-id="s903"><ol class="steps"><li value="14"><div class="step-editable-wrapper"><div class="step-editable multi-editable editable selectable is-locked" contenteditable="false" data-cid="s903" data-index="14">Elution fraction was dialyzed against 1L of Dialysis Buffer (30 mM Tris-HCl (pH 7.6), 10 mM EDTA) at 4C overnight. </div></div><div class="time-editable-wrapper"><div class="time-editable force-visible editable selectable is-locked" contenteditable="false" data-cid="time-s903" data-index="14" data-original-title="" title=""></div><span class="glyphicon glyphicon-time time-editable-wrapper-icon" icon="time"></span></div></li></ol></div></div></div></div></div></div></div></div></div></div><!-- react-empty: 154 --></div></div></div>
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